Targeting Microvesicles in Kidney Disease

项目总结 细胞外小泡(EV)运输和传递信号分子到远处的细胞以支持新陈代谢 过程、肾脏发育和动态平衡。EV信号的放松与新陈代谢和 慢性肾脏疾病。EV Cargo抑制葡萄糖转运蛋白，导致胰岛素抵抗。此外，电动汽车 刺激促纤维化因子以推动糖尿病患者的肾脏纤维化。此外，EV还牵涉到 多囊肾病的发病机制。尽管越来越多的人意识到EV在肾脏中扮演着至关重要的角色 疾病，我们对控制EV形成和货物装载的分子原理的基本知识仍然 从根本上说是有限的。 在预测性动物模型中识别和靶向控制EV生物发生的分子有可能 为肾脏疾病患者带来新的和协同的治疗方法。 EV的释放是一个进化保守的过程，从藻类到苍蝇再到人类，这表明从广义上讲 这一过程对分子的要求是可以概括的。果蝇在黑腹果蝇中具有无价的价值 发现与人类疾病相关的保守信号动力学，包括肾病。开始，开始 为了探索EV释放和载货的分子机制，我们建立了体内EV标记 系统在果蝇上皮组织中。该系统允许在活体成像中可视化EV分泌步骤 接近了。重要的是，我们已经将该系统用于EV蛋白质组研究，目的是识别新的 电动汽车生物生成和货物装载的调节器。此外，该系统还适用于目标基因突变 分析，允许对假定的电动汽车调节器进行可靠的功能验证。在这项试探性的R21拨款中，我们是 提议使用我们的系统来识别新的电动汽车调节器。经过验证的监管机构将构成遵循的基础- UP R01 GRANT专注于用组织培养和小鼠模型描绘潜在的机制 慢性肾脏疾病。

PROJECT SUMMARY Extracellular vesicles (EV) transport and deliver signaling molecules to distant cells to support metabolic processes, kidney development and homeostasis. Deregulation of EV signals are associated with metabolic and chronic kidney disease. EV cargo inhibit glucose transporters, leading to insulin resistance. In addition, EV stimulate profibrotic factors to drive kidney fibrosis in diabetic patients. Furthermore, EV are implicated in the pathogenesis of polycystic kidney disease. Despite the growing realization that EV play a vital role in kidney disease, our basic knowledge of the molecular principles that control EV formation and cargo loading remains fundamentally limited. Identifying and targeting molecules that control EV biogenesis in predictive animal models has the potential to lead to novel and synergizing therapeutics for the benefit of kidney disease patients. EV release is an evolutionary conserved process from algae to flies to humans, suggesting that broadly generalizable molecular requirements for this process exist. Drosophila melanogaster has been invaluable in the discovery of conserved signaling dynamics relevant to human diseases, including nephropathies. To begin to explore the molecular mechanism of EV release and cargo loading, we have developed an in vivo EV labeling system in Drosophila epithelial tissues. This system permits the visualization of EV secretion steps in live imaging approaches. Importantly, we have adapted this system for EV proteomic studies aimed at identifying novel regulators of EV biogenesis and cargo loading. Further, the system is amenable to targeted genetic mutant analyses, permitting robust functional validation of putative EV regulators. In this exploratory R21 grant, we are proposing to use our systems to identify novel EV regulators. Validated regulators will form the basis of a follow- up R01 grant focused on delineating the underlying mechanisms using tissue culture and murine models of chronic kidney disease.